title: A pH-sensitive gene delivery system based on folic acid-PEG-chitosan - PAMAM-plasmid DNA complexes for cancer cell targeting.
abstract: In this study, pH-sensitive biomaterials coated polymer/DNA nanocomplexes containing a high mobility group box 1 (HMGB1) were developed as an efficient non-viral gene delivery system. HMGB1 is a family of endogenous molecules that contains nuclear locating sequences (NSL). Polyethylene glycol tethered carboxylated chitosan modified with folic acid (FA-PEG-CCTS) was synthesized and its buffering capacity was determined by acid-base titration. A pH-sensitive core-shell system FA-PEG-CCTS/PAMAM/HMGB1/pDNA nanocomplexes (FPCPHDs), was prepared and characterized. Electrophoresis showed that FPCPHDs were resistant to heparin replacement and DNase I digestion. FPCPHDs exhibited only minor toxic effects on HepG2 and KB cells. The results of both luciferase activity assay and RFP fluorescence intensity analysis showed that FPCPHDs enhanced gene transfection and expression in KB cells. Moreover, gene transfection and expression in KB cells were inhibited by free folic acid. Intracellular trafficking of FPCPHDs in KB cells showed that FPCPHDs could rapidly escape from endo-lysosomes and become exclusively located in the nucleus at 3 h post transfection. In addition, FPCPHDs exhibited increased red fluorescence protein (RFP) expression at the tumor site of S180 xenograft nude mice. All results suggest that FPCPHDs is an efficient approach to improve the transfection and expression efficiency in most FR-positive cancer cells.
authors.affiliation: Department of Pharmaceutics, School of Pharmacy, Shenyang Pharmaceutical University, No. 103, Wenhua Road, Shenyang 110016, PR China.
authors.forename: Mingyue
authors.initials: M
authors.lastname: Wang
authors.forename: Haiyang
authors.initials: H
authors.lastname: Hu
authors.forename: Yuqi
authors.initials: Y
authors.lastname: Sun
authors.forename: Lipeng
authors.initials: L
authors.lastname: Qiu
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Guannan
authors.initials: G
authors.lastname: Guan
authors.forename: Xiuli
authors.initials: X
authors.lastname: Zhao
authors.forename: Mingxi
authors.initials: M
authors.lastname: Qiao
authors.forename: Liang
authors.initials: L
authors.lastname: Cheng
authors.forename: Lifang
authors.initials: L
authors.lastname: Cheng
authors.forename: Dawei
authors.initials: D
authors.lastname: Chen
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D048271:Chitosan / Q000737:chemistry*; D004247:DNA / Q000008:administration & dosage / Q000235:genetics*; D050091:Dendrimers / Q000737:chemistry*; D005260:Female; D005492:Folic Acid / Q000737:chemistry*; D018014:Gene Transfer Techniques; D056945:Hep G2 Cells; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics; D011092:Polyethylene Glycols / Q000737:chemistry*
chemical_list: D050091:Dendrimers; C522249:PEG-PAMAM; D011092:Polyethylene Glycols; D004247:DNA; D048271:Chitosan; D005492:Folic Acid
keywords: Gene delivery system; Gene transfection; Lysosomal escape; Nuclear locating sequence; Tumor targeting